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. 2008 Feb 1;177(3):301-8.
doi: 10.1164/rccm.200703-515OC. Epub 2007 Nov 7.

CD40 and CD80/86 act synergistically to regulate inflammation and mortality in polymicrobial sepsis

Anna Nolan  1 , Michael Weiden, Ann Kelly, Yoshihiko Hoshino, Satomi Hoshino, Nehal Mehta, Jeffrey A Gold
Affiliations

CD40 and CD80/86 act synergistically to regulate inflammation and mortality in polymicrobial sepsis

Anna Nolan et al. Am J Respir Crit Care Med. .

Abstract

Rationale: Costimulatory molecules, including the CD40-CD154 and CD80/86-CD28 dyads, play a prominent role in regulating inflammation in the adaptive immune response. Studies from our group and others suggest a potentially important role for these costimulatory cascades in innate immunity as well.

Objectives: To determine the role of CD80/86 alone and in combination with CD40 in lethal polymicrobial sepsis in mice and humans.

Methods: The murine cecal ligation and puncture (CLP) model was used to determine the role of CD80/86 alone and in combination with CD40 using wild-type mice, CD80/86(-/-) mice, and novel CD40/80/86(-/-) mice. Expression of cell-bound and soluble costimulatory molecules was assessed in humans via ELISA and flow cytometry.

Measurements and main results: Lethal CLP was associated with up-regulation of CD40 and CD80/86 and their respective ligands CD28 and CD154 on innate effector cells. Blockade or deletion of CD80/86 attenuated mortality and inflammatory cytokine production during CLP. CD40/80/86(-/-) mice exhibited further reductions in mortality, lung injury, and inflammatory cytokine production compared with CD80/86(-/-) mice. Finally, humans with sepsis had increased monocyte expression of CD40 and CD80 compared with healthy control subjects; with higher levels in subjects requiring vasopressor support. Levels of soluble CD28 and CD154 were significantly higher in patients who died compared with those who lived.

Conclusions: These data demonstrate a central role for CD40 and CD80/86 in the innate immune response and suggest that combined inhibition of CD40 and CD80/86 may improve mortality in sepsis. Expression of costimulatory molecules may serve as biomarkers for outcome in septic patients.

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Figures

<b>Figure 1.</b>
Figure 1.
Expression of CD40 and CD86 system after cecal ligation and puncture (CLP). C57BL/6 mice underwent CLP with a 19-gauge needle and were killed 18 hours later for flow cytometric analysis. Representative histograms for control and septic mice and a composite graph are presented for each molecule. Gray-shaded areas represent isotype control. (A) CD80 expression on peritoneal lavage mononuclear (CD11b+) cells; P < 0.001. (B) CD86 expression on splenic mononuclear cells; P < 0.001. (C) CD28 expression on peripheral blood neutrophils (PMNs) (Ly6G+); P = 0.05. (D) CD154 expression on CD11b+Ly6G+ peripheral blood mononuclear cells; P = 0.04. (E) CD40 expression on CD11b+Ly6G+ peripheral blood mononuclear cells; P = 0.04. Data represent three to five mice per group and were analyzed by Mann-Whitney t test.
<b>Figure 2.</b>
Figure 2.
Inhibition of CD80/86 alone or in combination with CD40 improves survival after cecal ligation and puncture (CLP). (A) Wild-type (WT) mice were injected intraperitoneally with either control IgG or 200 μg each of α-CD80 and α-CD86 4 hours before CLP and monitored for survival. n = 15 mice per group. (B) WT (n = 18), CD40−/− (n = 10), CD80/86−/− (n = 18), and CD40/80/86−/− (n = 15) mice underwent 19-gauge CLP and were monitored for survival. Data from CD40−/− mice have been previously published (31). For all survival experiments mice were monitored for a total of 14 days. Data are presented for the first 7 days, as there was no additional attrition between Days 7 and 14.
<b>Figure 3.</b>
Figure 3.
CD40/80/86−/− mice have attenuated cytokine production after cecal ligation and puncture (CLP). Wild-type (WT), CD80/86−/−, and CD40/80/86−/− mice underwent 19-gauge CLP and were killed 18 hours after CLP. Plasma and bronchoalveolar lavage fluid (BALF) were collected and analyzed for IL-6 (A) IL-12 (B), and IL-10 (C) by commercially available ELISA. There were six to eight mice in each group. Data were analyzed by nonparametric analysis of variance, with P values shown at the bottom of each graph. Statistically significant group comparisons are bracketed.
<b>Figure 4.</b>
Figure 4.
CD40/80/86−/− mice have additional attenuation in remote organ injury after cecal ligation and puncture (CLP). CD40/80/86−/− mice have attenuated cytokine production after CLP. Wild-type (WT), CD80/86−/−, and CD40/80/86−/− mice underwent 19-gauge CLP and were killed 18 hours after CLP. (A) Pulmonary capillary leakage was determined with Evans blue dye as previously described (18, 31). (B) nuclear factor (NF)-κB was determined by p65 DNA–binding ELISA from liver homogenates. Data are normalized for total protein content. (C) Liver myeloperoxidase (MPO) was determined by ELISA. Data are presented as nanograms of MPO per microgram of hepatic protein. (D) Peripheral blood bacterial cultures 18 hours after CLP, as determined by serial dilutions. There were four to six mice per group. Data were analyzed by nonparametric analysis of variance, with P values shown at the bottom of each graph. Statistically significant group comparisons are bracketed.
<b>Figure 5.</b>
Figure 5.
Neutrophils (PMNs) induce IL-6 production in vivo via CD40 and CD80/86. PMNs (5 × 105) from wild-type (WT) mice were injected intraperitoneally into WT, CD80/86−/−, or CD40/80/86−/− mice for 4 hours, at which point (A) plasma and (B) peritoneal lavage (PL) fluid were collected and analyzed for IL-6 via ELISA. There were five to eight mice per group. Data were analyzed by nonparametric analysis of variance, with P values shown at the bottom of each graph. Statistically significant group comparisons are bracketed.
<b>Figure 6.</b>
Figure 6.
Humans with sepsis present alterations in costimulatory molecules. Healthy control subjects and 14 septic subjects underwent flow cytometric analysis for CD40 (A), CD80 (B), and CD86 (D) on circulating monocytes. Monocytes were identified on the basis of forward scatter/side scatter (FSC/SSC) and CD14+. Septic subjects had blood collected on Days 1, 3, 7, and 14 or until hospital discharge. Thirteen of 14 septic subjects survived until discharge. *P < 0.01 compared with control subjects. (C) Expression levels of CD40 on CD14+ monocytes in healthy control subjects (n = 6), septic subjects (n = 4), and septic shock patients (n = 10) on Day 1. Data were analyzed by nonparametric analysis of variance, with the P value shown at the bottom of the graph. Statistically significant group comparisons are bracketed.
<b>Figure 7.</b>
Figure 7.
Expression of soluble CD28 and CD154 correlates with lethality in human sepsis. Levels of soluble CD154 (sCD154; A) and soluble CD28 (sCD28; B) were measured by ELISA in healthy control subjects (n = 11) and septic subjects (n = 35) on Day 1. Levels for both were higher in nonsurvivors (n = 6) compared with survivors (n = 29). Data were analyzed by nonparametric analysis of variance, P < 0.001. Statistically significant group comparisons are bracketed.
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References

    1. Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J, Pinsky MR. Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care. Crit Care Med 2001;29:1303–1310. - PubMed
    1. Hotchkiss RS, Karl IE. The pathophysiology and treatment of sepsis. N Engl J Med 2003;348:138–150. - PubMed
    1. Opal SM, Fisher CJ Jr, Dhainaut JF, Vincent JL, Brase R, Lowry SF, Sadoff JC, Slotman GJ, Levy H, Balk RA, et al.; Interleukin-1 Receptor Antagonist Sepsis Investigator Group. Confirmatory interleukin-1 receptor antagonist trial in severe sepsis: a phase III, randomized, double-blind, placebo-controlled, multicenter trial. Crit Care Med 1997;25:1115–1124. - PubMed
    1. Abraham E, Anzueto A, Gutierrez G, Tessler S, San Pedro G, Wunderink R, Dal Nogare A, Nasraway S, Berman S, Cooney R, et al.; NORASEPT II Study Group. Double-blind randomised controlled trial of monoclonal antibody to human tumour necrosis factor in treatment of septic shock. Lancet 1998;351:929–933. - PubMed
    1. Cohen J, Carlet J; International Sepsis Trial Study Group. INTERSEPT: an international, multicenter, placebo-controlled trial of monoclonal antibody to human tumor necrosis factor-α in patients with sepsis. Crit Care Med 1996;24:1431–1440. - PubMed

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